Serum long-noncoding RNA H19 and beta-catenin as biomarkers for early diagnosis of colorectal cancer in egyptian patients: a case control study

Colorectal cancer (CRC) is the third most prevalent cancer and the second most common cause of cancer death; however, its early detection can improve the survival. Colonic polyps are considered one of the CRC's major risk factors. Throughout many biological processes and malignancies, the non-coding RNAs have essential functions. Certain long noncoding RNAs (lncRNAs), including H19, were supposed to be CRC possible biomarkers. Also, H19 has been reported to play a role in regulating the activity of beta-catenin, a protein that regulates cell-to-cell adhesion, as well as gene transcription. The current work aimed to investigate the potential significance of LncRNA H19 relative serum expression level by quantitative polymerase chain reaction (q-PCR) and beta-catenin by enzyme-linked immunosorbent assay (ELISA) as noninvasive biomarkers to discriminate between colorectal cancer and colonic polyps. The statistical analysis of the studied factors revealed that the serum expression of H19 and beta-catenin in cancer cases were substantially greater than colonic polyp cases and normal control. Conclusion: The relative expressions of H19 and beta-catenin in the serum can significantly discriminate patients with CRC from those with polyp and normal controls, which could help when screening for CRC. (AU)

MicroRNA 30a Mediated Autophagy and Imatinib Response in Egyptian Chronic Myeloid Leukemia Patients.

Imatinib Mesylate is the drug used for targeted tyrosine kinase inhibition in the beginning of management of all Chronic Myeloid Leukemia (CML) newly diagnosed cases. However, resistance presents a considerable limit to its efficacy. Currently, it is impossible to anticipate IM resistance which makes the recognition of early flags an important treatment goal in CML. In this work we studied the connection between microRNA 30a (miR-30a) and Beclin 1 mediated autophagy and IM resistance in Egyptian CML patients. The study included newly diagnosed (group I, n = 20), imatinib responder (group II, n = 30), imatinib resistant (group III, n = 30) CML patients and a healthy demographically matched control group (group IV, n = 20). miR-30a expression was assayed by quantitative reverse transcription polymerase chain reaction. The variation in expression of miR-30a between CML cases and healthy controls was calculated using relative quantification method (2-ΔΔCT). Beclin 1 was assayed in Peripheral blood mononuclear cells by western blotting. miR-30a was over expressed and Beclin 1 was under expressed in imatinib responders compared to resistant cases median 1.21(0.55-3.02) versus median 0.65 (0.03-1.0) (p = 0.001) and median 950.0 (400.0-2410.0) versus, median 1570.0 (920.0-5430.0) (p < 0.001) respectively. Beclin 1 correlated significantly positively with miR-30a in new cases (p = 0.001) and negatively in imatinib responders (p = 0.021). Receiver Operating Curves demonstrated the performances of miR-30a and Beclin 1 to detect imatinib resistance. They showed sensitivities of 97.14% and 94.29% and specificities of 53.33% and 42.22% at the cut-off values of 1 and 940 respectively. Both miR-30a and Beclin 1 levels showed a relation with imatinib response and can therefore be put forward as valuable markers for detection of resistance and may also have promising future therapeutic implications.

The effectiveness of assertiveness training program on psychological wellbeing and work engagement among novice psychiatric nurses.

AIM: The study aimed to assess the effectiveness of an assertiveness training program on psychological wellbeing and work engagement among novice psychiatric nurses. DESIGN: A quasi-experimental design was utilized (one group pre/post assessment). SETTING: The study was conducted at The Abbasia hospital for mental health in Cairo, Egypt. SUBJECTS: The subjects of the study were 36 novice nurses who were starting their careers the hospital during 2017/2018. TOOLS: The current study used four tools for collecting the data: socio-demographic data sheet, Rathus Assertiveness Schedule, Riff's Psychological Well-Being Scale, and Utrecht Work Engagement Scale. RESULTS: The study results revealed a statistically significant difference between measure one and measure two intervention program regarding assertiveness skills, psychological well-being, and work engagement. Also, there was a significant positive correlation between the total mean scores of assertiveness skills and total mean scores of psychological well-being. CONCLUSIONS: This single-group feasibility study demonstrated that assertiveness training for novice nurses seems feasible. It may achieve a favorable outcome in developing assertiveness skills and improving psychological wellbeing. RECOMMENDATIONS: Further randomized controlled trials with more extended follow-up periods are required.

Evaluation of possible molecular toxicity induced by occupational exposure to lead and concomitant effect of smoking.

One of the most toxic heavy metals in the environment nowadays is lead (Pb). Even though exposure to lead has been reduced in some developed countries, individuals working in certain occupations are still exposed to lead at dangerous levels. Occupational exposure is of great concern and is also the main cause of lead poisoning. Although experts in various fields have been investigating the toxic effects of lead and its compounds for many years now, the association between chronic lead exposure and geno-toxicity is still an interesting point of research. The study aims to evaluate the possible DNA damage and the oxidative stress status induced by occupational exposure to lead and the role of concomitant smoking. The study was conducted on 60 subjects divided into two groups: an exposed group (40 male workers exposed to lead in their workplaces). This group was further divided into two subgroups; 20 workers were cigarette smokers and the other 20 workers were non-smokers. The other control group consists of 20 healthy males, not exposed to lead and matched by age to the exposed group (10 were smokers and the rest were non-smokers). Venous blood samples were collected from each participant for the determination of the following: blood lead level (BLL), plasma malondialdehyde (MDA) levels, and DNA damage using agarose gel electrophoresis. The exposed workers had significantly higher levels of lead and MDA, as well as a high frequency of DNA fragmentation. Smoking workers showed a greater frequency of DNA fragmentation than non-smokers. A significant relation was revealed between the BLL, as well as the MDA level, and the degree of DNA fragmentation among the lead-exposed workers. The study has shown additional evidence proving the association between Pb exposure and oxidative stress. The results further reinforced the role of cigarette smoking in augmenting such oxidative damage in the Pb-exposed population. However, further studies are recommended to evaluate the effect of cigarette smoking on Pb-exposed workers.

Combinatorial strategy of epigenetic and hormonal therapies: A novel promising approach for treating advanced prostate cancer.

AIMS: Estrogens act as key factors in prostate biology, cellular proliferation and differentiation as well as cancer development and progression. The expression of estrogen receptor (ER)-ß appears to be lost during prostate cancer progression through hypermethylation mechanism. Epigenetic drugs such as 5-aza-2'-deoxycytidine (5-AZAC) and Trichostatin A (TSA) showed efficacy in restoring ERß expression in prostate cancer cells. This study was designed to explore the potential anti-carcinogenic effects resulting from re-expressing ERß1 using 5-AZAC and/or TSA, followed by its stimulation with Diarylpropionitrile (DPN), a selective ERß1 agonist, in prostate cancer cell line PC-3. MAIN METHODS: Cells were treated with 5-AZAC, TSA, DPN and their combination. Subsequently, they were subjected to proliferation assays, determinations of ERß1 expression, protein levels of active caspase-3, cyclin D1, ß-catenin and VEGF. KEY FINDINGS: Treatment with these drugs exhibited an increase in ERß1 expression to different extents as well as active caspase-3 levels. Meanwhile, a significant reduction in cyclin D1, VEGF and ß-catenin levels was achieved as compared to the vehicle control group (p < 0.05). Interestingly, the triple combination regimen led to the most prominent anti-tumor responses in terms of increased apoptosis, reduced proliferation as well as angiogenesis. SIGNIFICANCE: The results support the notion that ERß1 acts as a tumor suppressor protein and suggest that sequential ERß1 expression and activation can offer significant anti-tumor responses. The study highlights that the strategy of merging epigenetic and hormonal therapies may be beneficial in treating advanced prostate cancer.

Total protein composition of young and adult Biomphalaria alexandrina snails with different compatibilities to Schistosoma mansoni infection / Composición total de proteína en caracoles Biomphalaria alexandrina juveniles y adultos con distintas compatibilidades a la infección por Schistosoma mansoni

Abstract:Schistosomiasis remains a disease of major global public health concern since it is a chronic and debilitating illness. The widely distributed Schistosoma mansoni that causes intestinal schistosomiasis represents a great threat. Its world-wide distribution is permitted by the broad geographic range of the susceptible species of its intermediate host, Biomphalaria, which serves as an obligatory host for the larval stage, at which humans get infected. The objectives were to identify the proteins responsible for the snails' compatibility outcome through differentiation between the total proteins among Biomphalaria alexandrina snails at different ages. The work was conducted on snails that differ in age and genetic backgrounds. Four subgroups (F1) from the progeny of self-reproduced susceptible and resistant snails (F0) were studied. Infection rates of these subgroups (young susceptible, adult susceptible, young resistant and adult resistant) were 90 %, 75 %, 40 % and 0 %, respectively. Using Sodium Dodecyl Sulphate Polyacrylamide Gel electrophoresis (SDS-PAGE), differences in protein expression were evaluated between adult and young snails of different subgroups. Dice similarity coefficient was calculated to determine the percentage of band sharing among the experimental subgroups. The results showed that the combination of similarities between age and compatibility status of the snails, lead to the highest similarity coefficient, followed by the combination of similarities of both genetic origin and age, even though they differ in the compatibility status. On the other hand, the differences in the genetic background, age and compatibility status, lead to the least similarity index. It was also found that the genetic background in young snails plays a major role in the determination of their compatibility, while the internal defense system has the upper hand in determining the level of adult compatibility. In conclusion, the findings of the present work highlight the great impact of the snail age in concomitance with the genetics and the internal defense in the determination of B. alexandrina/S.mansoni compatibility. Future works are recommended, as further characterization of the shared protein bands among the studied subgroups is needed to clarify their role in host-parasite relationship. Rev. Biol. Trop. 64 (4): 1747-1757. Epub 2016 December 01.

Resumen:La esquistosomiasis es una enfermedad crónica y debilitante que constituye una problemática de salud pública a nivel mundial. Debido a que Schistosoma mansoni está ampliamente distribuida y a que es el causante de la esquistosomiasis intestinal representa una gran amenaza. Biomphalaria es el hospedero intermedio y obligatorio para el estado larval, presenta una amplia distribución geográfica e infecta al ser humano. El objetivo fue identificar las proteínas responsables del efecto de compatibilidad en caracoles Biomphalaria alexandrina de distintos estadios a través de la diferenciación del total de proteínas en ellos. La investigación se llevó a cabo con caracoles de diferentes edades y antecedentes genéticos. Se estudiaron cuatro subgrupos (F1) de la progenie de caracoles susceptibles y resistentes reproducidos asexualmente (F0). Las tasas de infección de estos subgrupos (juvenil susceptible, adulto susceptible, juvenil resistente, adulto resistente) fueron 90 %, 75 %, 40 % y 0 %, respectivamente. Con la electroforesis en gel de poliacrilamida en presencia de dodecilsulfato sódico (SDS-PAGE) se evaluaron las diferencias en la expresión proteica entre los caracoles juveniles y adultos de los distintos subgrupos. Se calculó el coeficiente de similitud de Dice para determinar el porcentaje de bandas compartidas entre los subgrupos experimentales. Los resultados mostraron que la combinación de similitudes entre la edad y el estado de compatibilidad de los caracoles genera el mayor coeficiente de similitud seguido por el de la combinación de similitudes tanto de la edad como del origen genético aunque varían en el estado de compatibilidad. Por otra parte, las diferencias en los antecedentes genéticos, la edad y el estado de compatibilidad generan el índice de similitud más bajo. También se encontró que el antecedente genético en caracoles juveniles es importante en la determinación de su compatibilidad, mientras que el sistema de defensa interno es el que determina el nivel de compatibilidad en adultos. En conclusión, los resultados de este trabajo resaltan la importancia de la edad del caracol en conjunto con la genética y la defensa interna para determinar la compatibilidad de B. alexandrina/S.mansoni. Se recomienda realizar futuros trabajos así como una mayor caracterización de las bandas proteicas compartidas entre los subgrupos estudiados para esclarecer su papel en la relación hospedero-parásito.

Total protein composition of young and adult Biomphalaria alexandrina snails with different compatibilities to Schistosoma mansoni infection.

Schistosomiasis remains a disease of major global public health concern since it is a chronic and debilitating illness. The widely distributed Schistosoma mansoni that causes intestinal schistosomiasis represents a great threat. Its world-wide distribution is permitted by the broad geographic range of the susceptible species of its intermediate host, Biomphalaria, which serves as an obligatory host for the larval stage, at which humans get infected. The objectives were to identify the proteins responsible for the snails' compatibility outcome through differentiation between the total proteins among Biomphalaria alexandrina snails at different ages. The work was conducted on snails that differ in age and genetic backgrounds. Four subgroups (F1) from the progeny of self-reproduced susceptible and resistant snails (F0) were studied. Infection rates of these subgroups (young susceptible, adult susceptible, young resistant and adult resistant) were 90 %, 75 %, 40 % and 0 %, respectively. Using Sodium Dodecyl Sulphate Polyacrylamide Gel electrophoresis (SDS-PAGE), differences in protein expression were evaluated between adult and young snails of different subgroups. Dice similarity coefficient was calculated to determine the percentage of band sharing among the experimental subgroups. The results showed that the combination of similarities between age and compatibility status of the snails, lead to the highest similarity coefficient, followed by the combination of similarities of both genetic origin and age, even though they differ in the compatibility status. On the other hand, the differences in the genetic background, age and compatibility status, lead to the least similarity index. It was also found that the genetic background in young snails plays a major role in the determination of their compatibility, while the internal defense system has the upper hand in determining the level of adult compatibility. In conclusion, the findings of the present work highlight the great impact of the snail age in concomitance with the genetics and the internal defense in the determination of B. alexandrina/S.mansoni compatibility. Future works are recommended, as further characterization of the shared protein bands among the studied subgroups is needed to clarify their role in host-parasite relationship.

Impact of the age of Biomphalaria alexandrina snails on Schistosoma mansoni transmission: modulation of the genetic outcome and the internal defence system of the snail

Of the approximately 34 identified Biomphalariaspecies,Biomphalaria alexandrinarepresents the intermediate host of Schistosoma mansoniin Egypt. Using parasitological and SOD1 enzyme assay, this study aimed to elucidate the impact of the age of B. alexandrinasnails on their genetic variability and internal defence against S. mansoniinfection. Susceptible and resistant snails were reared individually for self-reproduction; four subgroups of their progeny were used in experiment. The young susceptible subgroup showed the highest infection rate, the shortest pre-patent period, the highest total cercarial production, the highest mortality rate and the lowest SOD1 activity. Among the young and adult susceptible subgroups, 8% and 26% were found to be resistant, indicating the inheritance of resistance alleles from parents. The adult resistant subgroup, however, contained only resistant snails and showed the highest enzyme activity. The complex interaction between snail age, genetic background and internal defence resulted in great variability in compatibility patterns, with the highest significant difference between young susceptible and adult resistant snails. The results demonstrate that resistance alleles function to a greater degree in adults, with higher SOD1 activity and provide potential implications for Biomphalariacontrol. The identification of the most susceptible snail age enables determination of the best timing for applying molluscicides. Moreover, adult resistant snails could be beneficial in biological snail control.

Impact of the age of Biomphalaria alexandrina snails on Schistosoma mansoni transmission: modulation of the genetic outcome and the internal defence system of the snail.

Of the approximately 34 identified Biomphalariaspecies,Biomphalaria alexandrinarepresents the intermediate host of Schistosoma mansoniin Egypt. Using parasitological and SOD1 enzyme assay, this study aimed to elucidate the impact of the age of B. alexandrinasnails on their genetic variability and internal defence against S. mansoniinfection. Susceptible and resistant snails were reared individually for self-reproduction; four subgroups of their progeny were used in experiment. The young susceptible subgroup showed the highest infection rate, the shortest pre-patent period, the highest total cercarial production, the highest mortality rate and the lowest SOD1 activity. Among the young and adult susceptible subgroups, 8% and 26% were found to be resistant, indicating the inheritance of resistance alleles from parents. The adult resistant subgroup, however, contained only resistant snails and showed the highest enzyme activity. The complex interaction between snail age, genetic background and internal defence resulted in great variability in compatibility patterns, with the highest significant difference between young susceptible and adult resistant snails. The results demonstrate that resistance alleles function to a greater degree in adults, with higher SOD1 activity and provide potential implications for Biomphalariacontrol. The identification of the most susceptible snail age enables determination of the best timing for applying molluscicides. Moreover, adult resistant snails could be beneficial in biological snail control.

Behavioral and neuronal biochemical possible effects in experimental induced chronic mild stress in male albino rats under the effect of oral barley administration in comparison to venlafaxine.

Venlafaxine is an antidepressant of choice, whose effectiveness could be modified by a commonly used medicinal plant and nutrient. The current study had evaluated the barley extract (1 g/kg) when compared to or combined to venlafaxine (32 mg/kg) in a rat stress model. The present study was conducted on 40 male Wister albino rats; divided to five groups. Four groups were subjected to social chronic mild stress. Drugs or saline were orally daily administered one week before stress induction and extended up to ten weeks. Behavioral, brain biochemical tests and serum magnesium were assessed at the end. The study revealed significant change in the combined group on behavioral tests; forced swim test, elevated plus maze and saccharin preference test when compared to barley extract group. Furthermore, there was significant reduction in brain malondialdehyde level, no significant change in brain nitric oxide level, while significant increase in serum magnesium level was noticed. Whereas, the barley extract group recorded a lowest significant improvement in behavioral, brain and serum biochemical tests. It could be concluded that barley and venlafaxine together had muffled the oxidant stress and increased brain serotonin, serum magnesium level that might had a crucial role in experimental induced chronic mild stress in rats.

Association of genetic polymorphism -670A>G in the Fas gene and serum markers AST platelet ratio index, AST/ALT with significant fibrosis and cirrhosis in chronic hepatitis C.

AIM: This study was carried out to evaluate the association of genetic polymorphism -670A>G in the promoter of Fas gene as well as serum biomarkers aspartate aminotransferase (AST) platelet ratio index (APRI) and AST/alanine aminotransferase (ALT) with significant fibrosis and cirrhosis in chronic hepatitis C patients. Seventy-nine patients with chronic hepatitis C in addition to 80 age- and sex-matched healthy controls were evaluated for genetic polymorphism -670A>G of Fas gene by polymerase chain reaction-restriction fragment length polymorphism and serum biomarkers APRI and AST/ALT in relation to significant fibrosis and cirrhosis diagnosed by liver biopsy. RESULT: Genetic polymorphism -670A>G in Fas gene was associated with significant liver fibrosis and cirrhosis. Heterozygous mutation was found in 11.4% of patients and 10% of controls, while homozygous mutation was found only in 7.6% of patients. Odds ratio (OR) was statistically not significant (OR=1.93, 95% confidence interval=0.76-4.92). Mean values of APRI and AST/ALT were significantly higher in patients with (F3-F4) compared with those with (F0-F2). (p-value <0.001 for APRI and p=0.007 for AST/ALT). In addition, APRI showed a better sensitivity than AST/ALT for prediction of significant fibrosis. CONCLUSION: Genetic polymorphism -670A>G of Fas gene was associated with significant fibrosis and cirrhosis in chronic hepatitis C patients. APRI and AST/ALT are independent predictors for significant fibrosis. APRI showed a better sensitivity than AST/ALT for prediction of significant fibrosis. Moreover, APRI can be used as an index to exclude liver cirrhosis without performing liver biopsy.

Experimental trichinosis: parasitological, electrophysiological and biochemical studies.

Trichinosis is a parasitic infection affecting the gut and the muscles causing mild gastrointestinal symptoms followed by periorbital oedema, muscle pains, fever and eosinophilia. The infection evokes functional disturbances in physiological effector systems. Furthermore, several biochemical changes are associated with the infection. Therefore, this work was carried out to study the electrophysiological changes in intestine, striated and cardiac muscles by electromyography (EMG) and to assess the biochemical changes through measurement of serum cholinesterase and intestinal myeloperoxidase activity (MPO) in both light and heavy infected experimental animals by Trichinella spiralis (T. spiralis). Electrophysiological results showed increased contractility of the smooth muscle layers of the intestine only early in the infection, whereas both striated and cardiac muscles showed increase in the contractility with the progress of infection in both light and heavy infection. Significant myocardial dysfunction in the form of bradycardia, in addition to major histopathological changes in the heart occurred from the beginning of the infection and increased till the end of the study. Biochemical study showed gradual increase in serum cholinesterase, while, the intestinal MPO showed increase only in the early stage of the infection. It was noticed that all changes were more pronounced in the heavily infected group than the lightly infected one.

Stat3 enhances vimentin gene expression by binding to the antisilencer element and interacting with the repressor protein, ZBP-89.

Vimentin exhibits a complex pattern of developmental- and tissue-specific expression and is aberrantly expressed in most metastatic tumors. The human vimentin promoter contains multiple DNA elements, some of which enhance gene expression and one that inhibits. A silencer element (at -319) binds the repressor ZBP-89. Further upstream (at -757) is an element, which acts positively in the presence of the silencer element and, thus, is referred to as an antisilencer (ASE). Previously, we showed that Stat1alpha binds to this element upon induction by IFN-gamma. However, substantial binding and reporter gene activity was still present in nontreated cells. Here, we have found that Stat3 binds to the ASE element in vitro. Transfection experiments in COS-1 cells with various vimentin promoter--reporter constructs show that gene activity is dependent upon the cotransfection and activation of Stat3. Moreover, activated Stat3 can overcome ZBP-89 repression. Coimmunoprecipitation studies demonstrate that Stat3 and ZBP-89 can interact and confocal microscopy detects these factors to be colocalized in the nucleus. Moreover, a correlation exists between the presence of activated Stat3 and vimentin expression in MDA-MB-231 cells, which is lacking in MCF7 cells where vimentin is not expressed. In the light of these results, we propose that the interaction of Stat3 and ZBP-89 may be crucial for overcoming the effects of the repressor ZBP-89, which suggests a novel mode for Stat3 gene activation.

ZBP-89 represses vimentin gene transcription by interacting with the transcriptional activator, Sp1.

Vimentin, a member of the intermediate filament protein family, is regulated both developmentally and tissue specifically. It is also a marker of the metastatic potential of many tumor cells. Pre viously, the human vimentin promoter has been shown to contain multiple elements for the binding of both positive- and negative-acting regulatory factors. Transient transfection analysis of various vimentin 5'-end promoter sequences and mutants thereof fused to a reporter gene further defined two regulatory elements, a positive element that binds Sp1 and a negative element that binds the protein ZBP-89. ZBP-89 has been shown to be either a repressor or an activator of gene expression, depending on the promoter. Here, we show that for vimentin, both ZBP-89 and ZBP-99 repress reporter gene expression in Schneider (S2) cells. Deletion constructs confirm that the glutamine-rich region of Sp1 is required to enhance vimentin transcription, whereas the N-terminus of ZBP-89 is required to interact with Sp1 and repress gene expression. The overexpression of hTAF(II)130 can alleviate ZBP-89 repression in S2 cells, suggesting how ZBP-89 might serve to block gene expression.